Simplified 3D protocol capable of generating early cortical neuroepithelium
Here, we report a 3D cerebellar differentiation protocol with quick startup method, defined medium and no special materials or handling requirements. Three fibroblast growth factors (FGF2, 4 and 8) were used for cerebellar patterning and smoothened agonist (SAG) for granule cell development. After 3...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
The Company of Biologists
2017-03-01
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| Series: | Biology Open |
| Subjects: | |
| Online Access: | http://bio.biologists.org/content/6/3/402 |
| _version_ | 1818678000387883008 |
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| author | Dwayne B. Holmes Vivi M. Heine |
| author_facet | Dwayne B. Holmes Vivi M. Heine |
| author_sort | Dwayne B. Holmes |
| collection | DOAJ |
| description | Here, we report a 3D cerebellar differentiation protocol with quick startup method, defined medium and no special materials or handling requirements. Three fibroblast growth factors (FGF2, 4 and 8) were used for cerebellar patterning and smoothened agonist (SAG) for granule cell development. After 35 days, differentiation products exhibited similar structures and neuronal markers reported in prior ‘organoid’ and ‘spheroid’ protocols. This included cells positive for KIRREL2 (a marker of early cerebellar neuroepithelium) and ZIC1 (a marker for granule cells). Follow-up tests indicated that addition of FGFs, if helpful, was not required to generate observed structures and cell types. This suggests that intrinsic production of patterning factors by aggregates themselves may be adequate for region-specific 3D modeling. This protocol may be used as a quick, easy and cost-efficient method for 3D culture, whether to research development of the early cerebellar neuroepithelium, a base to generate mature cortical structures, or to optimize minimal-factor protocols for other brain regions. |
| first_indexed | 2024-12-17T09:08:18Z |
| format | Article |
| id | doaj.art-ede6d239485248c8a66a697a819fc6f2 |
| institution | Directory Open Access Journal |
| issn | 2046-6390 |
| language | English |
| last_indexed | 2024-12-17T09:08:18Z |
| publishDate | 2017-03-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Biology Open |
| spelling | doaj.art-ede6d239485248c8a66a697a819fc6f22022-12-21T21:55:23ZengThe Company of BiologistsBiology Open2046-63902017-03-016340240610.1242/bio.021725021725Simplified 3D protocol capable of generating early cortical neuroepitheliumDwayne B. Holmes0Vivi M. Heine1 Department of Pediatrics/Child Neurology, Amsterdam Neuroscience, VU University Medical Center, Amsterdam 1081 HV, The Netherlands Department of Pediatrics/Child Neurology, Amsterdam Neuroscience, VU University Medical Center, Amsterdam 1081 HV, The Netherlands Here, we report a 3D cerebellar differentiation protocol with quick startup method, defined medium and no special materials or handling requirements. Three fibroblast growth factors (FGF2, 4 and 8) were used for cerebellar patterning and smoothened agonist (SAG) for granule cell development. After 35 days, differentiation products exhibited similar structures and neuronal markers reported in prior ‘organoid’ and ‘spheroid’ protocols. This included cells positive for KIRREL2 (a marker of early cerebellar neuroepithelium) and ZIC1 (a marker for granule cells). Follow-up tests indicated that addition of FGFs, if helpful, was not required to generate observed structures and cell types. This suggests that intrinsic production of patterning factors by aggregates themselves may be adequate for region-specific 3D modeling. This protocol may be used as a quick, easy and cost-efficient method for 3D culture, whether to research development of the early cerebellar neuroepithelium, a base to generate mature cortical structures, or to optimize minimal-factor protocols for other brain regions.http://bio.biologists.org/content/6/3/4023DOrganoidPluripotent stem cellCerebellumCortex |
| spellingShingle | Dwayne B. Holmes Vivi M. Heine Simplified 3D protocol capable of generating early cortical neuroepithelium Biology Open 3D Organoid Pluripotent stem cell Cerebellum Cortex |
| title | Simplified 3D protocol capable of generating early cortical neuroepithelium |
| title_full | Simplified 3D protocol capable of generating early cortical neuroepithelium |
| title_fullStr | Simplified 3D protocol capable of generating early cortical neuroepithelium |
| title_full_unstemmed | Simplified 3D protocol capable of generating early cortical neuroepithelium |
| title_short | Simplified 3D protocol capable of generating early cortical neuroepithelium |
| title_sort | simplified 3d protocol capable of generating early cortical neuroepithelium |
| topic | 3D Organoid Pluripotent stem cell Cerebellum Cortex |
| url | http://bio.biologists.org/content/6/3/402 |
| work_keys_str_mv | AT dwaynebholmes simplified3dprotocolcapableofgeneratingearlycorticalneuroepithelium AT vivimheine simplified3dprotocolcapableofgeneratingearlycorticalneuroepithelium |